Evolution of whole cells and organisms by recursive sequence recombination

A method of evolving a cell to acquire a desired property, comprising:

• (i.) forming protoplasts of a population of different cells;
• (ii.) fusing the protoplasts to form hybrid protoplasts, in which genomes from the protoplasts recombine to form hybrid genomes;
• (iii.) incubating the hybrid protoplasts under conditions promoting regeneration of cells, thereby producing regenerated cells;
• (iv.) repeatedly forming protoplasts from the regenerated cells, fusing the protoplasts to form hybrid protoplasts, in which genomes from the protoplasts recombine to form additional hybrid genomes;
• incubating the additional hybrid protoplasts under conditions promoting regeneration of cells, thereby producing additional regenerated cells;
• and,
• (v.) selecting or screening to isolate regenerated cells or additionally regenerated cells that have evolved toward acquisition of the desired property.

A method of poolwise recombination and selection of cells, the method comprising:

• (i) introducing a library of DNA fragments into a plurality of recipient cells, whereby a plurality of fragments of the library undergo recombination with a genome or an episome of a plurality of recipient cells to produce a plurality of modified cells, whereby the plurality of modified cells comprise a plurality of different modified fragments;
• (ii) fusing at least two of the modified cells, whereby genomes of the at least two modified cells recombine;
• (iii) segregating the resulting fused cell to produce a plurality of regenerated cells, each with a single genome;
• and,
• (iv) selecting the regenerated cells for a desired property.